Cloning and characterization of a human c-myc promoter-binding protein

Journal Article · Mon Apr 01 04:00:00 EST 1991 · Molecular and Cellular Biology; (United States)

DOI:https://doi.org/10.1128/MCB.11.4.2154· OSTI ID:7014245

Ray, R; Miller, D M ^[1]

---

1. University of Alabama at Birmingham, Alabama (United States)

A human cDNA clone encoding a c-myc promoter-binding protein was detected by screening a HeLa cell lambda phage expression cDNA library. The library was screened by using an XhoI-NaeI human c-myc P[sub 2] promoter fragment as a probe. The recombinant phage encoded a fusion protein, myc-binding protein 1 (MBP-1), which has an apparent molecular size of 40 kDa. A corresponding protein with a molecular size of 35 kDa was present in a HeLa cell extract. Sequence analysis of the cloned gene reveals an open reading frame of 1,038 bp with a 3[prime] untranslated region of 378 bp. The predicted protein sequence contains a proline-rich region in the amino terminus but does not demonstrate a known DNA-binding domain. DNase I footprint analysis demonstrates that MBP-1 binds to the sequence just 5[prime] of the TATA box sequence of the human c-myc P[sub 2] promoter. MBP-1 cDNA hybridizes to a 1.4-kb mRNA from HeLa and HL-60 cells, indicating that the cDNA insert (1,416 bp) is a full-length clone. Coexpression of the MBP-1 protein represses transcription from the human c-myc promoter, suggesting that MBP-1 may act as a negative regulatory factor for the human c-myc gene.

• OSTI ID: 7014245
• Journal Information: Molecular and Cellular Biology; (United States), Journal Name: Molecular and Cellular Biology; (United States) Vol. 11:4; ISSN 0270-7306; ISSN MCEBD4
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
BACTERIOPHAGES
CLONING
DNA
DNA HYBRIDIZATION
DNA-ASE
DNA-CLONING
ENZYMES
ESTERASES
GENE RECOMBINATION
GENES
HELA CELLS
HYBRIDIZATION
HYDROLASES
MICROORGANISMS
MOLECULAR BIOLOGY
NUCLEIC ACIDS
NUCLEOPROTEINS
ONCOGENES
ORGANIC COMPOUNDS
PARASITES
PHOSPHODIESTERASES
PROTEINS
RECOMBINANT DNA
VIRUSES